i bet 762  (Bio-Rad)

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: BRD4 alleviates ferroptosis induced by Erastin in a panel of five cell lines including HEK293T, HeLa, HepG2, RKO and PC3 cells. A Representative pictures and propidium iodide staining of HEK293T, HeLa, HepG2, RKO and PC3 cells after Erastin (20 μM) treatment for 24 h. Representative pictures and propidium iodide staining of ( B ) HEK293T cells, HeLa cells, ( C ) HepG2 cells, RKO cells and PC3 cells after being treated with DMSO, JQ-1 (1 μM), I-BET-762 (2 μM), Erastin (20 μM) or Erastin plus JQ-1, I-BET-762 respectively for 48 h. D CCK-8 was used to detect the cell viability of HEK293T cells and HeLa cells. E Representative pictures of HEK293T cells transfected with vector or plasmids encoding BRD4 after being treated with DMSO or Erastin (20 μM) respectively for 48 h. F Representative pictures of control and stable BRD4 knockdown cells in 293 T cells being treated with Erastin (20 μM) for 48 h. G Representative pictures of control and stable BRD4 knockdown cells in HeLa cells being treated with Erastin (20 μM) for 48 h. *p < 0 .05; **p < 0 .01; ***p < 0.001; ****p < 0.0001. Values are mean ± SD, n = 5

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Staining, CCK-8 Assay, Transfection, Plasmid Preparation

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on FTH1 and FTL in HEK293T cells and HeLa cells. HEK293T cells and HeLa cells were treated with JQ-1 and I-BET-762 for 24 h. A The protein levels of FTH1 and FTL in HEK293T cells were detected by Western blot. B qRT-PCR analysis of the mRNA expression levels of FTH1 and FTL in HEK293T cells. C The protein levels of FTH1 and FTL in HeLa cells were detected by Western blot. D qRT-PCR analysis of the mRNA expression levels of FTH1 and FTL in HeLa cells. *p < 0 .05; **p < 0 .01; ***p < 0.001

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Western Blot, Quantitative RT-PCR, Expressing

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on ROS levels in HEK293T cells and HeLa cells. HEK293T cells and HeLa cells were treated with JQ-1 (3 μM) and I-BET-762 (5 μM) respectively for 24 h. Rosup (50 μg/mL) was used as positive control to treat cells for 1 h. Reactive oxygen species assay kit was used to assess the ROS level in cells. Representative pictures of ROS level in HEK293T cells after being treated with ( A ) JQ and B I-BET-762. Representative pictures of ROS level in HeLa cells after being treated with ( C ) JQ and D I-BET-762. E Representative pictures of ROS level in control and stable BRD4 knockdown cells in 293 T cells. F Representative pictures of ROS level in control and stable BRD4 knockdown cells in HeLa cells. G 293T and Hela cells were treated with JQ-1 or I-BET-762. The level of malondialdehyde (MDA) was measured in these cells. H The level of malondialdehyde (MDA) was measured in BRD4 knocking down cells

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Positive Control

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on VDAC2 and VDAC3 in HEK293T cells and HeLa cells. HEK293T cells and HeLa cells were treated with JQ-1 and I-BET-762 for 24 h. A The protein level of VDAC2 in HEK293T cells was detected by Western blot. B qRT-PCR analysis of the mRNA expression levels of VDAC2 and VDAC3 in HEK293T cells. C The protein levels of VDAC2 in HeLa cells were detected by Western blot. D qRT-PCR analysis of the mRNA expression levels of VDAC2 and VDAC3 in HeLa cells. *p < 0 .05; **p < 0 .01; ***p < 0.001

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Western Blot, Quantitative RT-PCR, Expressing

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on Nrf2 in HEK293T cells and HeLa cells. A Western blot was used to detect the protein level of Nrf2 in HEK293T cells and HeLa cells with MG132 treatment. HEK293T cells and HeLa cells were treated with JQ-1 and I-BET-762 for 24 h. B NFR2 protein levels were measured by Western blot. NRF2 mRNA levels in HEK293T cells ( C ) and HeLa cells ( D ) were determined by qRT-PCR. **p < 0 .01; ***p < 0.001

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on System Xc and GPX4 in HEK293T cells and HeLa cells. HEK293T cells and HeLa cells were treated with JQ-1 and I-BET-762 for 24 h. A The protein level of GPX4 in HEK293T cells was detected by Western blot. B qRT-PCR analysis of the mRNA expression levels of SLC7A11, SLC3A2, and GPX4 in HEK293T cells. C Detected the protein levels of GPX4 in HeLa cells by Western blot. D The mRNA expression levels of SLC7A11, SLC3A2, and GPX4 in HeLa cells were measured by qRT-PCR. *p < 0 .05; **p < 0 .01; ***p < 0.001

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Western Blot, Quantitative RT-PCR, Expressing

Journal: Discover. Oncology

Article Title: BRD4 inhibitors broadly promote erastin-induced ferroptosis in different cell lines by targeting ROS and FSP1

doi: 10.1007/s12672-024-00928-y

Figure Lengend Snippet: Effect of BRD4 on the expression of FSP1 in HEK293T cells and HeLa cells. HEK293T cells and HeLa cells were treated with JQ-1 and I-BET-762 for 24 h. The protein level of FSP1 in ( A ) HEK293T cells and ( C ) HeLa cells were detected by Western blot. The mRNA expression levels of FSP1 in ( B ) HEK293T cells and ( D ) HeLa cells were measured by qRT-PCR. E The mRNA levels of FSP1 in control and stable BRD4 knockdown cells in HEK293T and HeLa cells were measured by qRT-PCR. F The protein levels of FSP1 in control and stable BRD4 knockdown cells in HEK293T and HeLa cells were measured by Western blot. G The ChIP sequencing data of BRD4 in SUM-159 cells with (SRX2194233) or without JQ-1(SRX2194281) treatment was download from GEO dataset and the data was analyzed by using USCS genome. HeLa cells were measured by qRT-PCR.*p < 0 .05; **p < 0 .01; ***p < 0.001

Article Snippet: The drugs include Erastin (B1524, APExBIO), JQ-1 (A1910, APExBIO), I-BET-762 (B1498, APExBIO), and MG132 (A2585, APExBIO).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, ChIP-sequencing